LOCUS       sequence-375784-        5893 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Single-integration vector for inserting PCP-NLS-GFPEnvy at the URA3 
            locus in Saccharomyces cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5893)
  AUTHORS   Brouwer I, Kerklingh E, van Leeuwen F, Lenstra TL
  TITLE     Dynamic epistasis analysis reveals how chromatin remodeling 
            regulates transcriptional bursting.
  JOURNAL   Nat Struct Mol Biol. 2023 May;30(5):692-702. doi: 
            10.1038/s41594-023-00981-1. Epub 2023 May 1.
  PUBMED    37127821
REFERENCE   2  (bases 1 to 5893)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41594-023-00981-1"; journalName: "Nat Struct Mol Biol"; 
            date: "2023-05"; volume: "30"; issue: "5"; pages: "692-702"
COMMENT     Sequence Label: SIV URA PCP-NLS-GFPEnvy
FEATURES             Location/Qualifiers
     source          1..5893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     256..273
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     257..273
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(302..318)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(302..318)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    319..352
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        389..732
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      1551..1748
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    1772..1805
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1806..1825
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1806..1824
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2846..2866
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      3611..3858
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3627..3645)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(3881..3899)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     rep_origin      complement(4187..4775)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4267..4286)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4946..5737,5738..5806))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5569..5588
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5807..5870)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taacgtgatg atatttcata aataatgtaa ttctatatat gttaattacc ttttttgcga
       61 ggcatattta tggtgaagga taagttttga ccatcaaaga aggttaatgt ggctgtggtt
      121 tcagggtcca taatgctttt caattcatct tttttttttt tgttcttttt tttgattccg
      181 gtttctttga aatttttttg attcggtaat ctccgagcag aaggaagaac gaaggaagga
      241 gcacagactt agatttgtaa aacgacggcc agtgcatgcg tatcggacgt ctcccttcat
      301 ggtcatagct gtttcctgat aacttcgtat aatgtatgct atacgaagtt attagcttgc
      361 ctcgtccccg ccgggtcacc cggcgagcga catggaggcc cagaataccc tccttgacag
      421 tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc ccgtacattt agcccataca
      481 tccccatgta taatcatttg catccataca ttttgatggc cgcacggcgc gaagcaaaaa
      541 ttacggctcc tcgctccaga cctgcgagca gggaaacgct cccctcacag acgcgttgaa
      601 ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt taggatttgc cactgaggtt
      661 cttctttcat atacttcctt ttaaaatctt gctaggatac agttctcaca tcacatccga
      721 acataaacaa ccatgacagt caacactaat acctatagtg agagagcaga aactcatgcc
      781 tcaccagtag cacagcgatt atttcgatta atggaactga agaaaaccaa tttatgtgca
      841 tcaattgacg ttgataccac taaggagttc cttgaattaa ttgataaatt aggtccttat
      901 gtatgcttaa tcaagactca tattgatata atcaatgatt tttcctatga atccactatt
      961 gaaccattat tagaactttc acgtaaacat caatttatga tttttgaaga tagaaaattt
     1021 gctgatattg gtaataccgt aaagaaacaa tatattggtg gagtttataa aattagtagt
     1081 tgggcagata ttaccaatgc tcatggtgtc actgggaatg gagtggttga aggattaaaa
     1141 cagggagcta aagaaaccac caccaaccaa gagccaagag ggttattgat gttagctgaa
     1201 ttatcatcag tgggatcatt agcatacgga gaatattctc aaaaaactgt tgaaattgct
     1261 aaatccgata aggaatttgt tattggattt attgcccaac gtgatatggg tggccaagaa
     1321 gaaggatttg attggcttat tatgacacct ggagttggat tagatgataa aggtgatgga
     1381 ttaggacaac aatatagaac tgttgatgaa gttgttagca ctggaactga tattatcatt
     1441 gttggtagag gattgtttgg taaaggaaga gatccagata ttgaaggtaa aaggtataga
     1501 aatgctggtt ggaatgctta tttgaaaaag actggccaat tataatcagt actgacaata
     1561 aaaagattct tgttttcaag aacttgtcat ttgtatagtt tttttatatt gtagttgttc
     1621 tattttaatc aaatgttagc gtgatttata ttttttttcg cctcgacatc atctgcccag
     1681 atgcgaagtt aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg
     1741 ctatactgct gtcgattcga tactaacgcc gataacttcg tataatgtat gctatacgaa
     1801 gttattaata cgactcacta tagggcgaat tggagctcgt attcaagata ttcttatcaa
     1861 agcacttatt ttttcattat atttctatat tgatttaagg acggtacgct attttctttg
     1921 taccttttac tatgcataaa aaaaggggaa gtgtttaaca atcgacactt acttaatatg
     1981 ttttgccgcc cttcataaga gggtgtttct aaaatttatt gggcaagaat gagatggact
     2041 cgcaccctac atgacgttta aatatttagt gttaaggttc agaacatgca ccaggtgcga
     2101 catgtgttgc gattatcatg acaatgtctc tatccgagat gcatttgtag tatcaattga
     2161 tgcgtattat gacatgattt acatagcata catcgtcaaa catgatatta tattcttttt
     2221 ttgataaatg tacggattta aagctgtcga atatattttc tgaaatttct tggagctgac
     2281 gcaaaatttt caaaggtgct aaaatttttc aagatttctc acttttgctt ggtaacaaag
     2341 aatgatggca ttgcattttt accaccggta catttaactg ctatttctca cgtttctttc
     2401 cctatcctta agtaattctt ttacaatcta agaaaaccac gatcaaacaa ataaatcagc
     2461 atctagaatg tccaaaacca tcgttctttc ggtcggcgag gctactcgca ctctgactga
     2521 gatccagtcc accgcagacc gtcagatctt cgaagagaag gtcgggcctc tggtgggtcg
     2581 gctgcgcctc acggcttcgc tccgtcaaaa cggagccaag accgcgtatc gcgtcaacct
     2641 aaaactggat caggcggacg tcgttgattc cggacttccg aaagtgcgct acactcaggt
     2701 atggtcgcac gacgtgacaa tcgttgcgaa tagcaccgag gcctcgcgca aatcgttgta
     2761 cgatttgacc aagtccctcg tcgcgacctc gcaggtcgaa gatcttgtcg tcaaccttgt
     2821 gccgctgggc cgtggatccc ccgggccaaa aaagaagaga aaggtagaat tcgatatcat
     2881 gtctaaaggc gaggaattgt ttacaggtgt agttcctatt ttggtcgaac tagatggaga
     2941 tgtcaacggt cataagtttt ctgtacgtgg tgaaggcgaa ggtgatgcta ctaatggcaa
     3001 actgactctg aagttcatct gtacgaccgg aaaattgcca gttccatggc ctacattagt
     3061 aacaacatta tgttacggag ttcaatgctt tgctagatac ccagatcaca tgaagcagca
     3121 tgactttttc aaaagtgcta tgccagaagg ttatgtgcag gagagaacta tctccttcaa
     3181 agatgatggt acttataaga caagagcaga ggtaaagttt gaaggcgata cattggttaa
     3241 tagaattgag cttaaaggca ttgattttaa ggaagatggg aatatcctcg gacataaact
     3301 tgaatacaat ttcaattcac ataatgttta tatcacagca gataaacaaa agaatggtat
     3361 aaaagcgaac tttaagacaa gacataacgt ggaagatggt agcgtccaat tagccgacca
     3421 ttaccaacaa aacactccta tcggggacgg gccagttctc cttccagaca accactacct
     3481 atcaacccaa tctgttctat ctaaagaccc taatgaaaaa cgagatcaca tggtcttgtt
     3541 agagttcgtg actgccgctg gtataacatt aggaatggac gaattgtaca aataaggatc
     3601 ctaactcgag tcatgtaatt agttatgtca cgcttacatt cacgccctcc ccccacatcc
     3661 gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt
     3721 atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac
     3781 agacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt tttgggacgc
     3841 tcgaaggctt taatttgcgg ccggtaccca gcttttgttc cctttagtga gggttaattt
     3901 cctgaaaact gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat
     3961 ttaattatat cagttattac gcgggaatct cggtcgtaat gatttctata atgacgaaaa
     4021 aaaaaaaatt ggaaagaaaa tgcttcatgg cctttataaa aaggaactat ccaatacctc
     4081 gccagaacca agtaacagta ttttacgggg cacaaatcaa gaacaataag acaggactgt
     4141 aaagatggac gcatttaatt aattcgaagg ccgcgttgct ggcgtttttc cataggctcc
     4201 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4261 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4321 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4381 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4441 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4501 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4561 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4621 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4681 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4741 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4801 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4861 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4921 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4981 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5041 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5101 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5161 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5221 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5281 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5341 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5401 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5461 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5521 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5581 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5641 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5701 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5761 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5821 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaagtat
     5881 cgttcgaatt aat
//
